Research Article
Volume 21 Issue 5 - 2022
Assessment and Analysis of Dental Pulp Stem Cells (DPSCs) Biomarkers and Viability Following Cryopreservation Reveals Novel Association with MiR-218 Expression

Keegan Lott1, Paris Collier1, Marc Ringor1, Austin Taylor2, Brennan Truman2, Katherine M Howard3 and Karl Kingsley3*

1School of Medicine, University of Nevada-Las Vegas, 2040 West Charleston Blvd, Las Vegas, Nevada, USA

2School of Dental Medicine, University of Nevada-Las Vegas, 1700 West Charleston Blvd, Las Vegas, Nevada, USA

3School of Dental Medicine, University of Nevada-Las Vegas, 1001 Shadow Lane, Las Vegas, Nevada, USA

*Corresponding Author: Karl Kingsley, School of Dental Medicine, University of Nevada-Las Vegas, 1001 Shadow Lane, Las Vegas, Nevada, USA.
Received: April 20, 2022; Published: April 29, 2022




Abstract

Introduction: Recent efforts have demonstrated the clinical use and application for dental-pulp derived stem cells or DPSCs, which have been demonstrated to function in many dental and non-dental, tissue-based applications. Despite these advances, few studies have evaluated the effects of cryopreservation However, as most dentally-derived DPSC are banked during orthodontic treatment in the adolescent and teenager years, recent studies have begun to evaluate the effects of long-term cryopreservation on DPSC for longer periods of time. The primary objective of this study was to determine the effects of long-term cryopreservation on DPSC viability and pluripotency over time intervals extending up to ten years.

Methods: This retrospective study was reviewed and approved by the UNLV Institutional Review Board (IRB). Using an existing DPSC repository, sixteen DPSC isolates were thawed, cultured and screened for viability. RNA was extracted from each cell line and screened for mesenchymal stem cell biomarkers and microRNA expression.

Results: Six (n = 6) rapidly doubling time (rDT), three (n = 3) intermediate (iDT), and seven (n = 7) slow (sDT) isolates were successfully thawed and cultured. Viability ranged from 17.6% to 49.3%, which did not exhibit a clear, linear association with length of cryopreservation or doubling time. Screening of RNA revealed all DPSC isolates expressed the mesenchymal stem cell biomarker Nestin, with variable expression of Sox-2, Oct-4 and NANOG also observed. However, microRNA screening revealed differential expression among the rDT (miR-21, miR-27), iDT and sDT (miR-124, miR-135, miR-218) DPSC isolates - with the highest viability observed among the miR-218 expressing DPSC cells.

Conclusions: Although many studies have focused on the potential for DPSC use in biomedical and bioengineering applications, few studies have evaluated the long-term effects of cryopreservation or methods for identifying which DPSC isolates may be most likely to survive this procedure. This study is among the first to evaluate DPSC cryopreservation for periods up to ten years, as well as to identify potential biomarkers (such as miR-218) that may identify DPSC isolates with high survival potential.

 

Keywords: Dental Pulp Stem Cells (DPSC); Cryopreservation; Biomarker; Microrna; QPCR

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Citation: Karl Kingsley., et al.“Assessment and Analysis of Dental Pulp Stem Cells (DPSCs) Biomarkers and Viability Following Cryopreservation Reveals Novel Association with MiR-218 Expression”. EC Dental Science 21.5 (2022): 115-128.

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